Bis quaternary oximes



United States Patent 2 Claims. (Cl. 167-65) (Granted under Title 35, US. Code (1952), sec. 266) This invention described herein may be manufactured and used by or for the Government of the United States of America for governmental purposes Without the payment to us of any noyalty thereon.

This is a division of application Serial No. 809,578,

filed April 28, 1959, now Patent No. 3,077,476, granted March 12, 1963.

This invention is directed to certain diquaternary pyridinium halide oximes which are useful as chemotherapeutic and prophylactic agents for mammals poisoned by 'anticholinesterases, particularly the nerve gas known as GB or sarin, i.e., isopropyl methylphosphohofiuoridate. V The invention relates to 1,1'-polymethylene bis (4- formylpyridinium) halide dioximes wherein the polymethylene group contains from 2 to 6 carbon atoms. These compounds have the structural formula N+ R U (3 OH=NOH OH=NOH. I where R is a polymethylene group having from two to six carbon atoms and X is chloride, bromide or iodide. This invention further includes compounds of Formula I, but in which R is a polymethylene group containing from seven to ten carbon atoms.

This invention further includes 1,l-(2,5-dimethyl-pphenylenedimethylene) bis =(4-formylpyridinium) halide dioximes of the formula L N\ OHP' -OHz N\ 2X .7 on,

CH=NOH CH=NOH 11 The invention also relates to 1,l' (2-butenylene) bis -(4-formylpyridinium) halide dioximes of the formula CH=NOH CH=N O H (III) The invention further relates to 1, 1'-polymethylene bis (3-formylpyridinium) halide dioximes of the formula 3,093,539 Patented June 11, 1963 dioximes of the formula I CH=NOH (IVb) This invention further relates to unsymmetrical bisquaternary 4-formylpyridinium halide monoximes of the formula CH=NOH (V) wherein R" represents either three lower alkyl groups or the hydrocarbon portion of the pyridine ring.

This invention also relates to the 1,1-trimethylene bis (3-amidooximopyridinium) halides The invention also relates to the compounds CH N 9a N 2X CH=N OH (VIII) While all these compounds are useful in varying degree for the purposes set out above, the different groups exhibit quite striking dififerences in effectiveness. In all the above formulas X* is chloride, bromide or iodide, which appear to be equivalents as to physiological action, except for their effect on solubility.

Wilson et al., in Patent No. 2,816,113, granted Dec. 10, 1957, disclose a group of compounds which are efiective as antidotes for treatment of'mammals poisoned by compounds of high anticholinesterase activity, such as the nerve gases diisopropylphosphorofluoridate -(DFP), isopropyl methylphosphonofluoridate (GB), and O-ethyl, N,N-dimethyt phosphoroamidocyanidate (GA), as well as other related organic phosphorus compounds, including many insecticides. The compound of the Wilson et a1. group which has received by far the most attention is 2- formyld-methyl pyridiniurn iodide oxime, commonly known as 2-pyridine aldoxime methiodide or 2-PAM. This compound is outstanding in its ability to reactivate, in vitro, acetyl cholinesterase which has been inhibited by, for example, GB. Thus, even as compared to the very closely related 4-PAM, disclosed in Example II of the Wilson patent, 2-PAM shows much greater activity. With isopropyl methylphosphonylated acetylcholinesterase the rate constant at pH 7.4 and 25 C. in the presence of 10* M acetylcholine is 2x10 per mole per minute for *2-PAM and 1.4 10 per mole per minute for 4-PAM. The Wilson patent shows a high rate of survival in mice which had been poisoned with paraoxon and then treated with Z-PAM.

Nerve gas poisoning has been treated symptomatically with drugs which are pharmacologically antagonistic to 3 acetyl choline. Such a compound is atropine and it is at present the recommended remedy. Recently, Z-PAM has been reported to enhance considerably the activity of atropine in the chemotherapeusis of poisoning due to organophosphorus compounds.

The compounds of Formula I above in which R contains from 2 to -6 carbon atoms are appreciably more effective than 2-PAM as reactivators of GB-inhibited acetylcholinesterase and also in enhancing the activity of atropine in both therapy and propylaxis. When X- is bromide, the variation of the rate constant for the in vitro reactivation of GB inhibited eel acetylcholinesterase at pH 7.4 and 25 C. was found to be as follows:

TABLE 1 R Rate constant (lJmoles/minutes) (CH2): 7x10 C 2 6x10 04 6X10" (CH2)5 1X104 (CH2)a 6Xl0 When administered in combination with atropine to animals poisoned with GB the order of effectiveness was somewhat different. Under these conditions the compound in which R=(CH i.e. 1,1'-trimethylene bis (4- f-ormylphyridinum) bromide dioxime also known as TMB-4, was most eifective. In rats challenged with a 2 LD dose of GB administered intravenously, all of a group of six animals survived if the atropine-TMB-4 combination was administered intravenously immediately after poisoning. The atropine-Z-PAM combination saved only two of the group of animals. On the other hand, with dogs which were given a 20 LD dose of GB subcutaneously the survival ratios were the same (4/5) for the two treatments, which were given intravenously when symptoms appeared. However, the recovery time was much shorter for the surviving animals which received the TMB-4, i.e., 2 hours, as against 24 hours for those receiving the Z-PAM.

A summary of the reactivation rates and survival ratios for these compounds when administered therapeutically to rats together with atropine is as follows:

TABLE 2 R Reactivation Survival rate constant ratio (GB) 7X10 6/6 6x10 6/6 6X10 e/e 1x10 6/6 6X10 3/4 These compounds constitute our presently preferred group.

Our compounds may be employed prophylactically, i.e., injected before exposure to the anticholinesterase agent, e.g., GB, or therapeutically, i.e., injected subsequent to exposure.

The following series of experiments compares the effectiveness of our presently preferred compound, TMB-4, with 2-PAM applied to various animals by these two Subcutaneous 0.900 mg./kg. LD

To minimize absorption effects both the GB and TMB- 4 were ordinarily given intravenously. However, in the therapeutic tests on rabbits, dogs and cats, the GB was administered subcutaneously, since death from 20 LD intravenous dose of GB occurs so quickly that it is virtually impossible to give timely administration of the antidote.

Atropine, when administered, was included in the following amounts.

Mg./kg. Rats 4 Rabbits I 2 Dogs and cats 0.5

The prophylactic doses were given within two minutes prior to the injection of the GB, the therapeutic doses so soon as poisoning symptoms were visible.

Table 3 shows the results.

TABLE 3 The recovery periods, i.e., time for disappearance of symptoms of poisoning, among survivors in the above tests, with atropine, were as follows.

TABLE 4 2PAM TMB-4 Animals Prophylactic Therapeutic Prophylactic Therapeutic Rats 60 min 15 min. Rabbits 3 hr 30 min 2 hr.

Cats 5 hr 5 hr 6 hr 24 hr.

D 24 hr 24 hr 1% hr 3 hr.

The compounds of Formula I in which R contains from 7 to 10 carbon atoms are less effective than those of our preferred group. For these compounds the reactivation rate constant and the survival ratio for rats (measured as given above) were as follows, X- being bromide.

TABLE 5 R Rate Survival Constant ratio (GB) 2x10 0/4 CH 1. 2x10 (CH2)10 1. 4X10 0/6 While these compounds were ineffective in vivo against GB, they were, together with 2-PAM, very eifective against certain other anticholinesterases, particularly that designated as VX by the US. Army Chemical Corps. All these compounds caused survival of all animals (survival rates of 4/4 and 6/6) when administered therapeutically to rats challenged by 2 LD doses of VX.

The compounds of Formula II exhibited properties intermediate those of the two subgroups of Formula I. When X- was chloride the compound had the following properties. (In this and all following tables the survival ratios are those for rats challenged by 2 LD doses of GB or (VX) and the oxime was employed therapeutically.)

Compounds of Formula III showed reactivation rates very close to those of our preferred group. Thus when X in Formula III is bromide the reactivation rate constant was 8X10 as compared to the value for the R==(CH member of our preferred group of 6x10 For the unsaturated member (Formula III) the survival ratio for rats challenged by GB was only A as compared to 6/ 6 for the saturated analogue (Formula I). Both gave complete survival (ratio of 4/4 and 6 6) for animals challenged by VX however.

Compounds of Formula I Va showed anomalous properties.

They gave reactivation rates which were low, but survival ratios which were high as compared to 2-PAM, as shown by Table 6, X- being bromide.

TABLE 6 R Reactivation Survival rate constant ratio (GB) (011m 3. 6X10 4/4 (CHM 4. 2X10 3/4 The compounds of group IVb, which are closely related to those of IVa, were somewhat less effective. When X" was bromide the compound had the following properties: Reactivation rate constant 2x10 survival ratio (GB)--2/4.

The compounds of group V were another group in which the results of therapeutic treatment against GB were better as compared to 2FPAM than the reactivation rate constants would suggest, as shown by the following table, X- being bromide.

The compounds of Formulas VI, VII and VIII, while being of difierent structure are alike in exhibiting reactivation rate constants which are very low as compared to 2- PAM but giving high survival ratios as shown by Table 8, X" being bromide in each case.

TABLE 8 Formula N0. Reactivation Survival rate ratio (GB) 69 4/4 Negligible 4/4 67 4/4 Preparation 0] Compounds 4-pyridinecarboxaldehyde oxime was prepared by warming on a steam bath a neutralized aqueous solution of 4- pyridine-carboxaldehyde and hydroxylamine hydrochlo- 6 ride. The oxime had a melting point of 130-130.5 C. The 2- and 3-oximes were produced by similar methods.

The quaternization to produce dioximes was carried out by reacting the proper oxime with a l, n dihaloalkane, 5 (CH X employing a 3:1 molar ratio of oxime to halide. The unsymmetrical quaternary monoximes were obtained by reacting the pyridine oxime with the appropriate omega-halopropyl quaternary salt in a 1.5:1 molar ratio. Two procedures were utilized.

Procedure A: A mixture of the pyridine oxime and halide was dissolved in sufficient ethanol and refluxed for the period of time specified in Table 9.

Procedure B: A mixture of the oxime and halide was dissolved in about 100 ml. of ethanol and heated in a 200 ml. capped pressure bottle (carbonated beverage type) for the length of time specified. The reaction mixtures were cooled to room temperature and the product removed by filtration. In several instances it was necessary to add absolute ether to effect complete precipitation. The products were recrystalized from ether. This procedure was usually employed because of its simplicity.

Table 9 gives the procedure, yields .and melting points for representative compounds.

TABLE 9 Substituents Melting (m) or decomposition (d) point, 0.

Yield percent H i-uaoooomq commence o: oeagwoooo gent-5x251:

Further details regarding the preparation and properties of certain of our compounds are given in the following publications, by us and our associates:

Pyridine Aldoximes by Edward J. Poziomek, Brennie E. Hackley, Jr., and George M. Steinberg, Journal of Organic Chemistry, vol. 23, pp. 714-717 (May 1958); and Chemotherapeutic Effectiveness of Trimethylene bis (4-Formyl Pyridinium Bromide) dioxime in Antichlolinesterase Poisoning, by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society for Experimental Biology and Medicine, vol. 98, pages 107-409 (May 1958). These articles are to be considered incorporated by reference in this specification.

While we have shown a number of specific examples of compounds and their use, it will be obvious that various changes can be made without departing from our invention, which is defined by the following claims.

We claim:

1. A method of therapeutically treating a mammal which has been poisoned by a compound having high anticholinesterase activity which comprises injecting a composition consisting of atropine and 1,1-(2,5 dimethyl-pphenylmethylene) bis-(4-formylpyridinium) halide dioxime.

2. A method of therapeutically treating a mammal which has been poisoned by a compound having high anticholinesterase activity which comprises injecting a composition consisting of atropine and 1,1'-(p-phenylene dimethylene) bis-(3-formyl pyridinium) halide dioxime.

References Cited in the file of this patent Poziomek, Amer. Chem. Soc. Abst. of Papers, 132nd Meeting, 1957, pp.l6-O. 

1. AMETHOD OF TERAPEUTICALLY TREATING A MAMMAL WHICH HAS BEEN POISONED BY A COMPOUND HAVING HIGH ANTICHOLINESTERASE ACTIVITY WHICH COMPRISES INJECTING A COMPOSITION CONSISTING OF ATROPINE AND 1,1''-(2,5 DIMETHYL-PPHENYLMETHYLENE) BIS-(4-FORMYLPYRIDINIUM) HALIDE DIOXME. 